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Agrobacterium Tumefaciens-Mediated Transformation of Strain of Penicillium Janthinellum GXCR

Author: YuRong
Tutor: LiYouZhi
School: Guangxi University
Course: Microbiology
Keywords: Micro purple the Penicillium strains of GXCR Heavy metal resistance mechanism Benomyl resistance Agrobacterium tumefaciens Genetic transformation
CLC: Q78
Type: Master's thesis
Year: 2008
Downloads: 224
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Abstract


Is of great significance to study the interaction between filamentous fungi and heavy metals from the genetic level , compared with the traditional method of genetic transformation in recent years, the development of Agrobacterium tumefaciens -mediated genetic transformation (ATMT) , ATMT with high efficiency , stability , the high - frequency single-copy random insertion , applicable to a wide range of filamentous fungi genetic transformation of one of the primary means of the present study . Based on the work of the laboratory has isolated a plant height of anti- variety of heavy metal salts and benomyl sensitive micro purple Penicillium (Penicillium janthinellum) strains GXCR good foundation , research will be established by the Agrobacterium tumefaciens LBA4404 mediated GXCR strain genetic transformation system for the study of the strains from the gene level , based on the possible mechanisms of resistance to heavy metal salts . The present study in plasmid pGSA1252 basis constructed T-DNA region containing benomyl resistance gene (Benomylresistance) PG -Bn binary vector is introduced into Agrobacterium tumefaciens LBA4404 by triparental transformation method , and diluted to 10 < benomyl resistant transformants were sup> 7 / ml of GXCR spores of training two days after the screening of 367 of the total DNA of 40 randomly picked transformants were PCR validation of 98% transformed strains can amplify the target fragment , the conversion efficiency of 3.7 transformed strains / 10 small bacteria block . Random selection of 30 transformant strains could be amplified target fragment , 13 transformant strains can not grow CuSO 4 PDA plates 9 the growth of the transformed strains was significantly affected by the of CuSO 4 < / sub> inhibition . Simultaneously the phenotype significantly change the total DNA of transformed strains by Southern hybridization analysis , the results show a conversion of 62.5% of the strains of the integration of a single copy of the T-DNA insert, easy analysis of the correlation function of a gene by insertion mutation , for the slightly purple the Penicillium strains GXCR the anti- of heavy metals genetic mechanism study valuable material .

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CLC: > Biological Sciences > Molecular Biology > Genetic engineering (genetic engineering)
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